Yuav ua li cas fluorescence microscopy txawv ntawm cov pa microscopy
Tsis ntev los no kuv tau sim ua qee qhov khov ntawm cov nas, thiab tam sim no kuv yuav tsum tau siv lub tshuab ntsuas fluorescence kom pom tias tus kab mob kuv txhaj yog nyob rau hauv lub hlwb xav tau. Qee lub hauv paus ntsiab lus ntawm fluorescence microscopy yuav tsum tau kawm luv luv, thiab kuv tseem yuav qhia rau lawv ntawm no.
Fluorescence microscope siv lub teeb ultraviolet los ua lub teeb ci los teeb pom cov khoom uas tau sim, ua rau cov khoom tawm ntawm lub teeb, thiab tom qab ntawd saib cov khoom hauv qab lub tshuab tsom. Feem ntau yog siv rau cov immunofluorescence hlwb, nws yog tsim los ntawm lub teeb qhov chaw, lim phaj system, thiab kho qhov muag system los soj ntsuam cov fluorescence duab ntawm tus qauv los ntawm magnification ntawm lub eyepiece thiab lub hom phiaj lens. Cia peb saib qhov sib txawv ntawm qhov no fluorescence microscope thiab lub tshuab kho qhov muag tsis tu ncua.
1. Hais txog txoj kev teeb pom kev zoo
Txoj kev teeb pom kev zoo ntawm lub tshuab hluav taws xob fluorescence feem ntau yog siv txoj kev poob qis, uas txhais tau hais tias lub teeb pom kev zoo tau muab tso rau hauv cov qauv ntsuas los ntawm lub hom phiaj lens.
2. Hais txog kev daws teeb meem
Fluorescence microscopy siv lub teeb ultraviolet ua lub teeb ci, nrog lub wavelength luv luv tab sis muaj kev daws teeb meem siab dua li cov khoom siv kho qhov muag zoo tib yam.
3. Qhov sib txawv ntawm lub lim
Lub fluorescence microscope siv ob lub lim dej tshwj xeeb, ib qho nyob rau pem hauv ntej ntawm lub teeb ci los lim tawm lub teeb pom kev, thiab lwm qhov ntawm lub hom phiaj thiab eyepiece los lim tawm lub teeb ultraviolet, uas tuaj yeem tiv thaiv lub qhov muag.
Fluorescence microscopy kuj yog ib hom ntawm optical microscopy, feem ntau vim hais tias lub wavelength zoo siab los ntawm fluorescence microscopy yog luv, uas ua rau muaj kev sib txawv ntawm cov qauv thiab siv cov fluorescence microscopy thiab zoo tib yam microscopy. Feem ntau cov fluorescence microscopes muaj kev ua haujlwm zoo ntawm kev ntes lub teeb tsis muaj zog, yog li lawv cov duab muaj peev xwm kuj zoo nyob rau hauv qhov tsis muaj zog fluorescence. Tsis tas li ntawd, nrog kev txhim kho txuas ntxiv ntawm fluorescence microscopy nyob rau xyoo tas los no, lub suab nrov kuj tau txo qis. Yog li ntawd, ntau thiab ntau dua fluorescence microscopes raug siv.
Kev paub txog ob-photon fluorescence microscopy
Lub hauv paus ntsiab lus ntawm ob-photon excitation yog tias nyob rau hauv siab photon ceev, fluorescent molecules ib txhij nqus ob lub wavelength photons, thiab tom qab ib tug luv luv lub sij hawm ntawm thiaj li hu ua zoo siab lub xeev lub neej, emit ib tug luv luv wavelength photon; Nws cov nyhuv yog tib yam li siv photon nrog lub wavelength ntawm ib nrab ntawm qhov ntev wavelength los ua kom cov fluorescent molecules. Ob lub photon excitation yuav tsum muaj photon ceev, thiab kom tsis txhob puas hlwb, lub zog siab hom-xauv mem tes laser yog siv nyob rau hauv ob-photon microscope. Lub laser emissions los ntawm hom laser no muaj lub siab ncov lub zog thiab qis nruab nrab lub zog, nrog ib tug mem tes dav ntawm tsuas yog 100 femtoseconds thiab ib tug zaus ntawm 80 mus rau 100 megahertz. Thaum siv lub hom phiaj ntawm cov lej aperture siab los tsom cov photons ntawm lub laser pulsed, photon ceev ntawm lub focal point ntawm lub hom phiaj yog siab tshaj, thiab ob-photon excitation tsuas yog tshwm sim ntawm lub focal point ntawm lub hom phiaj. Yog li ntawd, ob-photon microscope tsis xav tau confocal pinholes, uas txhim kho cov txiaj ntsig ntawm kev tshawb pom fluorescence.
Feem ntau fluorescence tshwm sim, vim qhov tsawg photon ceev ntawm excitation, ib tug fluorescent molecule tsuas nqus tau ib photon tib lub sij hawm, thiab ces emit ib fluorescent photon los ntawm hluav taws xob hloov, uas yog hu ua ib tug photon fluorescence. Rau cov txheej txheem fluorescence excitation siv laser ua lub teeb qhov chaw, ob-photon los yog multiphoton fluorescence phenomena yuav tshwm sim. Nyob rau lub sijhawm no, lub teeb pom kev zoo siv yog qhov siv siab, thiab qhov ntom ntom ntawm photon ua tau raws li qhov yuav tsum tau muaj rau cov fluorescent molecules kom nqus ob photons ib txhij. Nyob rau hauv tus txheej txheem ntawm kev siv ib tug raug laser raws li lub excitation teeb qhov chaw, lub photon ceev tseem tsis txaus los tsim ob-photon absorption tshwm sim. Feem ntau, femtosecond mem tes lasers siv, thiab lawv lub zog tam sim no tuaj yeem ncav cuag qib megawatts. Yog li ntawd, lub wavelength ntawm ob-photon fluorescence yog luv dua li excitation, uas yog sib npaug rau cov nyhuv tsim los ntawm ib nrab excitation wavelength excitation.
